Abstract: :
Purpose: To generate gene expression profiles of wildtype andNrl knockout mice at different developmental time-points andto identify differentially expressed genes. Methods: RNA wasisolated from both Nrl+/+ and Nrl-/- mice retina at four stagesof development, postnatal day 0 (P0), P2, P10 and P21. Fivemicrogram of total RNA, either from retina or a common referenceRNA source, was labeled with Cy5 or Cy3 dye respectively using3DNA® SubmicroTM Oligo Expression Array Detection Kit (Genisphere).These labeled targets were simultaneously hybridized to I-genemicroarray slides produced in the Kellogg Eye Center, Universityof Michigan. The hybridized slides were then washed, and scannedusing an Affymetrix 428 scanner to generate images of fluorescenceintensity, which were then converted into expression data byGleamsTM (NuTec). Five replicate experiments were performedfor each time-point to control for experimental variabilityand to enable statistical inference. Microarray data was thennormalized, analyzed and clustered using appropriate statisticalmodels. Results: Gene expression profiles for both Nrl+/+ andNrl-/- mice retina during four critical developmental ages havebeen generated relative to a common reference RNA. A comparisonbetween Nrl-/- and Nrl+/+ mice at P21 has identified a numberof differentially expressed genes such as S-opsin, Rhodopsin,Gnat1 and several novel transcripts. Further analysis and resultswill be presented. Conclusion: Microarray analysis has beenutilized to generate retinal gene expression profiles in wildtypeand Nrl knockout mice. Up-regulated genes in Nrl-/- mice maycontribute to cone development and function, while down-regulatedgenes may involve in rod development. These expression profileswill also assist in the identification of novel retinal diseasegenes and the study of cellular pathways of photoreceptor developmentand cone survival.